Isolation of Human Monoclonal Antibodies That Neutralize Human Rotavirus

Higo-Moriguchi, Kyoko; Akahori, Yasushi; Iba, Yoshitaka; Kurosawa, Yoshikazu; Taniguchi, Koki

doi:10.1128/jvi.78.10.5534.2004

Cited by 10 publications

(20 citation statements)

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“…Conversion of Fab-B4 to rhIgG1 or rhIgG4 was performed as previously described (27), with some modifications. The V region and C region L chain (V L C L ) and the V region H chain (V H ) of the Fab-B4 fragment were amplified by PCR with primers containing a restriction site as follows: V L C L , forward primer, 59-CTACTCTGGCTC-CGCGGTGCCAGACAGTCTGTGTTGACGCAGCGG-39, reverse primer, 59-TCGACTGGCGCGCCCTATGAACATTCTGTAGGGGCCACTGTCT-TC-39; V H , forward primer, 59-TTCCTCCTACTAGTGGCAGCTCCCA-GATGGGTCCTGTCCCAGGTGCAGCT GGTGCAGTCTGG-39, reverse primer, 59-GGTGGAGGCACTCGAGACGGTGACCGTGGTCC-39.…”

Section: Passive Transfer Studiesmentioning

confidence: 99%

Bullous Pemphigoid Autoantibodies Directly Induce Blister Formation without Complement Activation

Ujiie

Sasaoka

Izumi

et al. 2014

The Journal of Immunology

107

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Complement activation and subsequent recruitment of inflammatory cells at the dermal/epidermal junction are thought to be essential for blister formation in bullous pemphigoid (BP), an autoimmune blistering disease induced by autoantibodies against type XVII collagen (COL17); however, this theory does not fully explain the pathological features of BP. Recently, the involvement of complement-independent pathways has been proposed. To directly address the question of the necessity of the complement activation in blister formation, we generated C3-deficient COL17-humanized mice. First, we show that passive transfer of autoantibodies from BP patients induced blister formation in neonatal C3-deficient COL17-humanized mice without complement activation. By using newly generated human and murine mAbs against the pathogenic noncollagenous 16A domain of COL17 with high (human IgG1, murine IgG2), low (murine IgG1), or no (human IgG4) complement activation abilities, we demonstrate that the deposition of Abs, and not complements, is relevant to the induction of blister formation in neonatal and adult mice. Notably, passive transfer of BP autoantibodies reduced the amount of COL17 in lesional mice skin, as observed in cultured normal human keratinocytes treated with the same Abs. Moreover, the COL17 depletion was associated with a ubiquitin/proteasome pathway. In conclusion, the COL17 depletion induced by BP autoantibodies, and not complement activation, is essential for the blister formation under our experimental system.

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Section: Passive Transfer Studiesmentioning

confidence: 99%

Bullous Pemphigoid Autoantibodies Directly Induce Blister Formation without Complement Activation

Ujiie

Sasaoka

Izumi

et al. 2014

The Journal of Immunology

107

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“…We isolated human monoclonal antibodies (MAb) to gH using an antibody library called AIMS4 constructed from B-lymphocyte-rich tissues of several dozen people (6,19). Nine clones were selected for their neutralizing ability and their Fab sequences of heavy (H) and light (L) chains and used, in addition to TI-57, an anti-gH human MAb from a hybridoma, to characterize the neutralization epitopes of gH (18).…”

Section: Varicella-zoster Virus (Vzv) Glycoprotein H (Gh)mentioning

confidence: 99%

Characterization of Neutralizing Epitopes of Varicella-Zoster Virus Glycoprotein H

Akahori

Suzuki

Daikoku

et al. 2009

J Virol

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Varicella-zoster virus (VZV) glycoprotein H (gH)is the major neutralization target of VZV, and its neutralizing epitope is conformational. Ten neutralizing human monoclonal antibodies to gH were used to map the epitopes by immunohistochemical analysis and were categorized into seven epitope groups. The combinational neutralization efficacy of two epitope groups was not synergistic. Each epitope was partially or completely resistant to concanavalin A blocking of the glycomoiety of gH, and their antibodies inhibited the cell-to-cell spread of infection. The neutralization epitope comprised at least seven independent protein portions of gH that served as the target to inhibit cell-to-cell spread.Varicella-zoster virus (VZV) glycoprotein H (gH) is the major target for neutralization (4,5,7,9,18), and it plays an important role in viral entry and cell-to-cell spread of infection (1,3,11,12,15). We isolated human monoclonal antibodies (MAb) to gH using an antibody library called AIMS4 constructed from B-lymphocyte-rich tissues of several dozen people (6,19). Nine clones were selected for their neutralizing ability and their Fab sequences of heavy (H) and light (L) chains and used, in addition to TI-57, an anti-gH human MAb from a hybridoma, to characterize the neutralization epitopes of gH (18). Our system makes it possible to use the Fab form, which has about one-third the molecular weight of immunoglobulin G (IgG), in order to eliminate the spatial interaction between the Fc or other unreacted Fab of IgG molecules on one gH molecule. The neutralizing epitopes of gH are conformational, making gH hardly detectable by Western blot or enzyme-linked immunosorbent assay, and therefore, the conformational epitopes were mapped immunohistochemically. The combinational neutralizing activity between two species of Fab protein A (Fabpp) forms and the inhibition of cell-to-cell infection were characterized, and the neutralization domain of gH was found to comprise a cluster of the seven neutralization epitopes and to prevent cell-to-cell infection.Human embryonic lung cells were used to propagate Oka varicella vaccine, and cell-free virus was obtained by sonication of infected cells in SPGC medium (phosphate-buffered saline [PBS] containing 0.1% sodium glutamate, 5% sucrose, and 10% fetal bovine serum) followed by centrifugation (13,14,16).Except for TI-57, each MAb was expressed in two forms: Fab-pp and Fab with an avidin tag (Fab-Avi-tag). Fab-pp corresponds to an Fab molecule fused with two domains of the Fc-binding protein A from Staphylococcus aureus (8) and purified on an IgG-conjugated column (19). Fab-Avi-tag is composed of an Fab bearing a 23-amino-acid-long peptide tag that can be biotinylated by the bacterial BirA biotin ligase (1). Fab-Avi-tag antibodies were purified by using SoftLink soft release avidin resin (Promega, Madison, WI).To map the neutralizing epitope by Fab-pp, VZV-infected cells in 24-well plates were fixed by air-drying and then with 50% methanol and 50% acetone. The Fab-pp form (5 g/ml in 0.5 ml of ...

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“…Three human neutralizing MAbs against rotavirus have been derived from a phage display library of B-lymphocyte cDNA from naturally infected humans (21). The phage antibodies were selected for binding to rotavirus strain KU virions, tested for neutralization of strain KU, and converted to immunoglobulin G1 MAbs through recombinant DNA manipulation.…”

Section: Vol 80 2006mentioning

confidence: 99%

High-Resolution Molecular and Antigen Structure of the VP8* Core of a Sialic Acid-Independent Human Rotavirus Strain

Monnier

Higo-Moriguchi

Sun

et al. 2006

J Virol

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The most intensively studied rotavirus strains initially attach to cells when the "heads" of their protruding spikes bind cell surface sialic acid. Rotavirus strains that cause disease in humans do not bind this ligand. The structure of the sialic acid binding head (the VP8* core) from the simian rotavirus strain RRV has been reported, and neutralization epitopes have been mapped onto its surface. We report here a 1.6-Å resolution crystal structure of the equivalent domain from the sialic acid-independent rotavirus strain DS-1, which causes gastroenteritis in humans. Although the RRV and DS-1 VP8* cores differ functionally, they share the same galectin-like fold. Differences between the RRV and DS-1 VP8* cores in the region that corresponds to the RRV sialic acid binding site make it unlikely that DS-1 VP8* binds an alternative carbohydrate ligand in this location. In the crystals, a surface cleft on each DS-1 VP8* core binds N-terminal residues from a neighboring molecule. This cleft may function as a ligand binding site during rotavirus replication. We also report an escape mutant analysis, which allows the mapping of heterotypic neutralizing epitopes recognized by human monoclonal antibodies onto the surface of the VP8* core. The distribution of escape mutations on the DS-1 VP8* core indicates that neutralizing antibodies that recognize VP8* of human rotavirus strains may bind a conformation of the spike that differs from those observed to date.

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Isolation of Human Monoclonal Antibodies That Neutralize Human Rotavirus

Cited by 10 publications

References 0 publications

Bullous Pemphigoid Autoantibodies Directly Induce Blister Formation without Complement Activation

Bullous Pemphigoid Autoantibodies Directly Induce Blister Formation without Complement Activation

Characterization of Neutralizing Epitopes of Varicella-Zoster Virus Glycoprotein H

High-Resolution Molecular and Antigen Structure of the VP8* Core of a Sialic Acid-Independent Human Rotavirus Strain

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